"recycling" Qiagen plasmid prep columns

John Bradsher jnbradsh at STUD.MED.CORNELL.EDU
Mon Sep 18 19:26:14 EST 1995


Dear netters:
        I have a general interest that I wonder if someone can help me
with.  I have used Qiagen plasmid prep columns for large scale
purifications, and I wonder if these things can be re-used.  I am by
training a biochemist and I would believe that one should be able to
extensively wash a column and regenerate it such that a subsequent round of
plasmid purification could be done.  Of course, each such column that is
used is identified with respect to the plasmid purified, such that
subsequent columns would only be used for purifications of the same
plasmid.  So, what I'm looking for here is empirical data on what things
people have tried, and what degree of success these methods have had.
Specific interests of mine would include what elutions that you have used
to eliminate the retained protein and RNA from these columns (including any
notes you may want to throw in on theory), what methods you prefer to
document the completeness of the elution of such garbage, and what kinds of
yields you have documented from subsequent plasmid preps.  This would also
include any data on the distribution of the product into supercoiled and
nicked circular species.  Additionally, I would like to know anything about
the stability of these columns in aqueous media, both before and after
regeneration, if such data is available.  To be to the point, documentation
counts here; anyone that can show me a good result from such a procedure
should also be able to give quantitative data on the success of the
process.  Let me know what you think.   Thanks





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